Figure 3

Introduction of selected siRNAs in vivo protected mice from FAS agonistic antibody (JO2)-induced death. (a) Seven-day survival (%) of NIH Swiss mice pretreated with siRNAs before administration of FAS agonistic Ab (JO2). siRNAs (3.5 mg/kg) targeting selected genes DEGS1, HNRNPL, WASF1, and APCS were complexed with Invivofectamine 2.0 Reagent and delivered by tail vein injection to livers of NIH Swiss mice (10 per group). LUC siRNA was used as a negative control. Seventy-two hours after siRNA introduction, mice were injected i.p. with JO2 (240 μg/kg) and survival was monitored for 7 days. P-values for differences in the proportion of mice surviving on day 7 post JO2 administration were calculated using Fisher’s Exact Test (two-tailed) (P=0.01 for WASF siRNA, P=0.03 for all other tested siRNAs). (b) Effectiveness of siRNAs targeting WASF, APCS, HNRNPL and DEGS in knocking down gene expression. RT–PCR of total liver RNA was used to determine the percentage of initial gene expression (with control siRNA targeting luciferase set at 100%) remaining in mice injected with gene-specific siRNAs. NIH Swiss mice (n=10 mice/group) were treated as in a and liver RNA was prepared from surviving mice on day 7 after JO2 injection. RT–PCR results were quantified using GelQuant.NET software provided by biochemlabsolutions.com. P-values were calculated using Student’s t-test. Error bars indicate S.D